Two efficient chalcone fluorescent probes (probe-KCN1 and probe-KCN2) were developed for the detection of thiophenols. Upon gradual addition of thiophenols to the fluorescent probes, the fluorescence intensity of the emission band at 550 nm is enhanced about 40-fold, with a large Stokes shift (130 nm). Probe-KCN1 responds to thiophenols with a good range of linearity and a detection limit of 79 nM (R-2 = 0.9915), and Probe-KCN2 responds selectively to thiophenols over other amino acids, common metal ions and other potential interferents with a detection limit of 96 nM (R-2 = 0.9978). The low-toxicity probe has been successfully used to detect thiophenols in samples of seawater. These results demonstrate that Probe-KCN is a class of specific probes that might provide a simple way to monitor changes in thiophenols at low concentrations in seawater samples.